Ns-2-nonenoate (HNEAcid) [65,66], a major pathway of HNE detoxification. In particular, in rat and in human brain, HNEAcid formation occurs in the mitochondria by ALDH5A [65]. The detoxification of these Title Loaded From File aldehydes is important for neurodegenerative disorders such as AD, since high levels of unsaturated lipid content increase brain vulnerability to oxidative damage [67]. Therefore up-regulation of ALDH5 could be protective against cell damage. Previous research identified ALDH4 as a p53-inducible gene [68]. The current study is the first to show that the lack of p53 increases ALDH5 expression levels, adding an additional p53-target gene. An increasing body of evidence places p53 as a member of an intriguing network that includes tumor 18334597 suppression and aging [69]. The tumor suppressor activity of p53 protects against malignant transformation but also enhances the aging process [69]. As the role of p53 in aging is still unclear, we designed this study gain insights into the role of p53 in the brain and its involvement in neuronal cell death. In conclusion, we identified brain mitochondrial proteins in p53 null mice that display crucial p53-dependent cellular functions in the central nervous system. Therefore, our results reinforce the concept that the lack of p53 could disturb cell homeostasis causing cells to stimulate defensive pathways. We also elaborated on the link between p53 and mitochondria as we used for the study mitochondria from p53 knock out mice. Since mitochondrial dysfunction is a key feature of neurodegenerative diseases such as AD, p53 conceivably could be a novel therapeutic target for the treatment of these disorders (Fig. 2).Author ContributionsConceived and designed the experiments: DAB. Performed the experiments: AF RS EB GC JC JBK. Analyzed the data: AF RS EB GS MP CM DSC DAB. Contributed reagents/materials/analysis tools: DAB JC JBK. Wrote the paper: AF RS DSC DAB.
Fluorescence-activated cell sorting (FACS) of live cells is typically performed using antibodies that bind to proteins present on the cell surface or using intracellular co-expressed fluorescent Title Loaded From File reporter proteins. For characterization of embryonic stem cells and induced pluripotent stem cells, the expression of transcription factors, such as Oct4, Nanog and Sox2, is at present the most meaningful indication of stemness, yet the presence of these intracellular proteins cannot be detected in living cells without the use of co-expressed fluorescent reporters [1,2,3]. It would be useful be able to omit the insertion of fluorescence reporters and instead sort based on transcription factor expression in a manner that is independent of genetic modification. The use of molecular beacons (MBs) as reporters for the presence of mRNA presents a method to sort stem cell populations based on their mRNA expression levels. All expressed mRNAs can be potential targets and thus could be used as sorting parameters. MBs consist of sequences of 25?0 bases in length with a fluorophore attached to the 59-end and a quencher molecule to the 39-end [4]. At 37uC, the MB forms a hairpin structure that causes the fluorophore to be quenched. The sequence within the loop of the hairpin is designed to be complementary to the target mRNA of interest (Figure 1A). Upon hybridization of the central loop of the MB to its target, the hairpin opens, correspondingly releasing the fluorophore from the quencher. Therefore, the MB reports only upon binding with the target mRNA.To explore the use of.Ns-2-nonenoate (HNEAcid) [65,66], a major pathway of HNE detoxification. In particular, in rat and in human brain, HNEAcid formation occurs in the mitochondria by ALDH5A [65]. The detoxification of these aldehydes is important for neurodegenerative disorders such as AD, since high levels of unsaturated lipid content increase brain vulnerability to oxidative damage [67]. Therefore up-regulation of ALDH5 could be protective against cell damage. Previous research identified ALDH4 as a p53-inducible gene [68]. The current study is the first to show that the lack of p53 increases ALDH5 expression levels, adding an additional p53-target gene. An increasing body of evidence places p53 as a member of an intriguing network that includes tumor 18334597 suppression and aging [69]. The tumor suppressor activity of p53 protects against malignant transformation but also enhances the aging process [69]. As the role of p53 in aging is still unclear, we designed this study gain insights into the role of p53 in the brain and its involvement in neuronal cell death. In conclusion, we identified brain mitochondrial proteins in p53 null mice that display crucial p53-dependent cellular functions in the central nervous system. Therefore, our results reinforce the concept that the lack of p53 could disturb cell homeostasis causing cells to stimulate defensive pathways. We also elaborated on the link between p53 and mitochondria as we used for the study mitochondria from p53 knock out mice. Since mitochondrial dysfunction is a key feature of neurodegenerative diseases such as AD, p53 conceivably could be a novel therapeutic target for the treatment of these disorders (Fig. 2).Author ContributionsConceived and designed the experiments: DAB. Performed the experiments: AF RS EB GC JC JBK. Analyzed the data: AF RS EB GS MP CM DSC DAB. Contributed reagents/materials/analysis tools: DAB JC JBK. Wrote the paper: AF RS DSC DAB.
Fluorescence-activated cell sorting (FACS) of live cells is typically performed using antibodies that bind to proteins present on the cell surface or using intracellular co-expressed fluorescent reporter proteins. For characterization of embryonic stem cells and induced pluripotent stem cells, the expression of transcription factors, such as Oct4, Nanog and Sox2, is at present the most meaningful indication of stemness, yet the presence of these intracellular proteins cannot be detected in living cells without the use of co-expressed fluorescent reporters [1,2,3]. It would be useful be able to omit the insertion of fluorescence reporters and instead sort based on transcription factor expression in a manner that is independent of genetic modification. The use of molecular beacons (MBs) as reporters for the presence of mRNA presents a method to sort stem cell populations based on their mRNA expression levels. All expressed mRNAs can be potential targets and thus could be used as sorting parameters. MBs consist of sequences of 25?0 bases in length with a fluorophore attached to the 59-end and a quencher molecule to the 39-end [4]. At 37uC, the MB forms a hairpin structure that causes the fluorophore to be quenched. The sequence within the loop of the hairpin is designed to be complementary to the target mRNA of interest (Figure 1A). Upon hybridization of the central loop of the MB to its target, the hairpin opens, correspondingly releasing the fluorophore from the quencher. Therefore, the MB reports only upon binding with the target mRNA.To explore the use of.